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. 2022 Nov 9;50(20):11600–11618. doi: 10.1093/nar/gkac948

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Proteins recruited to persistently stalled replication forks. (A) Volcano plot displaying the results from n = 2 biologically independent SILAC iPOND experiments after 18 h hydroxyurea (HU) treatment against untreated control. FDR was calculated using limma. Selected proteins with an FDR < 5% are highlighted. (B) GO term analysis (Molecular Function, Biological Process and Cellular Compartment) of proteins enriched at persistently stalled replication forks (18 h HU) with an FDR < 5%. P-values were calculated by Fisher′s exact test and corrected for multiple comparisons using Benjamini–Hochberg correction. (C) Western blot validation of indicated proteins enriched by iPOND after 2 or 18 h hydroxyurea (HU) treatment. (D) Representative box plot of a proximity ligation assay (PLA) of n = 2 biologically independent experiments between TPX2 and RPA2 after treatment with DMSO or hydroxyurea (HU) for 2 or 18 h. Individual measurements of the mean PLA intensity per nucleus are plotted and the median indicated in red. P-values (****P-value < 0.0001) were derived from using one-way-ANOVA with Tukey correction for multiple comparisons. (E) Representative box plot of a proximity ligation assay (PLA) of n = 2 biologically independent experiments between PARP1 and RPA2 after treatment with DMSO or hydroxyurea (HU) for 2 or 18 h. Individual measurements of the mean PLA intensity per nucleus are plotted and the median indicated in red. P-values (****P-value < 0.0001) were derived from using one-way-ANOVA with Tukey correction for multiple comparisons.